The objectives of this proposal are to design, prepare, and test, new polymeric separation media in spherical bead form for use as packing materials in high performance liquid chromatography. The new media could complement, and in some cases replace, existing silica-based materials while introducing more "separation chemistry" into the media themselves. In view of the relative lack of stability of the current silica-based column packings to certain experimental conditions during chromatographic analyses or column regeneration, particular areas where improvement are sought include the following: (1) increased stability to severe elution conditions such as pH gradients in aqueous media; (2) improved compatibility with the biological activity of some proteins; (3) ability to separate a wide array of polar molecular including acidic or basic compounds such as are found in drugs, metabolites, and natural products; (4) increased efficiency over existing polymeric packing materials. Meeting these objectives will be accomplished by developing new techniques which will produce small (1-10 um) and preferably monodispersed crosslinked by polymer particles having a spherical shape, as well as incorporating new reactive groups within the polymers for enhanced interactions with the mixtures to be separated. In addition to developing new monomers and crosslinkers incorporating "separation chemistry", research will be carried out in the production of small diameter polymeric particles by suspension, dispersion, or seeded polymerizations. Both porous and non-porous particles will be studied. In the case of porous particles, conditions which produce high surface areas, appropriate pore-size distributions, and mechanical stability, will be optimized. In the case of non-porous particles, the use of light, high energy radiation, or in situ polymerization, to stabilize the particles by crosslinking will receive particular attention. In all cases, special attention will be paid to the surface chemistry of the polymer particles to enhance their chromatographic properties. Finally, actual chromatographic testing of the polymer beads with model compounds as well as actual biological materials will take place with optimization of the best materials.